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Solon Thanos

Publications and source records attributed to Solon Thanos.

At least 19 recordsLinked to original sources
PubMed2007

Transformation of adult retina from the regenerative to the axonogenesis state activates specific genes in various subsets of neurons and glial cells.

The purpose of this study was to identify the gene expression profile of the regenerating retina in vitro. To achieve this goal, three experimental groups were studied: (1) an injury control group (OC-LI group) that underwent open crush (OC) of the optic nerve and lens injury (LI) in vivo; (2) an experimental group (OC-LI-R group) that comprised animals treated like those in the OC-LI group except that retinal axons were allowed to regenerate (R) in vitro; and (3) an experimental group (OC-LI-NR group) that comprised animals treated as those in the OC-LI group, except that the retinas were cultured in vitro with the retinal ganglion cell (RGC) layer facing upwards to prevent axonal regeneration (NR). Gene expression in each treatment group was compared to that of untreated controls. Immunohistochemistry was used to examine whether expression of differentially regulated genes also occurred at the protein level and to localize these proteins to the respective retinal cells. Genes that were regulated belonged to different functional categories such as antioxidants, antiapoptotic molecules, transcription factors, secreted signaling molecules, inflammation-related genes, and others. Comparison of changes in gene expression among the various treatment groups revealed a relatively small cohort of genes that was expressed in different subsets of cells only in the OC-LI-R group; these genes can be considered to be regeneration-specific. Our findings demonstrate that axonal regeneration of RGC involves an orchestrated response of all retinal neurons and glia, and could provide a platform for the development of therapeutic strategies for the regeneration of injured ganglion cells.

Animals↗
PubMed2006

Pathological changes in human retinal ganglion cells associated with diabetic and hypertensive retinopathy.

BACKGROUND: To examine whether systemic diseases like diabetes and arterial hypertension, which frequently cause retinopathies leading to blindness effect the morphology of retinal ganglion cells (RGC). METHODS: Histological retina material with a history of being untreated, or laser-coagulated (LC) diabetic retinopathy (DR), or arterial hypertensive retinopathy (AHR) was used. The RGC were labeled by introducing crystals of the fluorescent carbocyanine dye DiI into the nerve fiber layer, which contains ganglion cell axons. RESULTS: The typical silhouettes of both major types of RGC, parasol and midget cells, were identified. The axons in DR and AHR retinas showed morphology changes such as irregular swelling and beading. Dendritic field sizes were significantly reduced in RGC of both the hypertonic and diabetic retinas. A significant reduction in branching frequency was evident in both the diabetic and hypertonic retinas, in both the midget and the parasol cells. In LC retinas, both parasol and midget RGC were observed within the LC spots, although their numbers were dramatically decreased compared with normal retinas. CONCLUSIONS: The data suggest that diabetes and arterial hypertonia have similar effects on the morphology of RGC, in addition to causing microvascular alterations and bleeding. Therefore, therapeutic measures and prognostic outcomes in diabetic and hypertensive retinopathy should also consider regressive changes in retinal neurons.

Adult↗
PubMed2006

Retinal gene profiling in a hereditary rodent model of elevated intraocular pressure.

PURPOSE: To characterize the changes in retinal gene expression induced by elevated intraocular pressure (IOP) in a hereditary rodent model. METHODS: A rat model derived from the RCS-rdy- strain develops IOP elevation spontaneously without experimental manipulation. Retinal gene expression after IOP elevation was compared with age-matched RCS-rdy- retinas having normal IOP levels The MWG Rat 10k array, which comprises 9715 rat genes spotted onto one array was used. Quantitative real-time PCR (qRT-PCR) was used to verify the expression of heat shock protein-27 (Hsp-27), SA hypertension-associated gene, c-myc, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), vascular endothelial growth factor (VEGF), myocilin, interleukin-7 (IL-7), mitogen activated protein kinase 13 (MAPK-13) and crystallin beta-A1 (Cryba1). The cellular distribution of c-myc, glial fibrillary acidic protein (GFAP), VEGF, and SA was assessed using immunohistochemistry. RESULTS: Elevated IOP of 37.7+/-5.0 mmHg shifted the retina's program of gene expression, with 75 genes being upregulated (equal to or higher than 3.0 fold) and 45 genes being downregulated (equal to or lower than 0.3 fold). These genes mediate various cellular processes such as cell adhesion, cell structure, hypertension, immunity, protein sythesis, proteolysis, transcription, and signaling. The regulation pattern of SA, VEGF, c-myc, IL-7, and MAPK-13, which are uniquely regulated in our model were confirmed by qRT-PCR experiments. The regulation of Hsp-27, TIMP-1, myocilin, and Cryba1, which have previously been associated with elevated IOP were also confirmed with qRT-PCR. The protein products of c-myc, SA, and GFAP were localized to astrocytes and Müller cells. Neurons in the ganglion cell layer and inner nuclear layer were VEGF-immunopositive. CONCLUSIONS: This study identified some of the genes that are differentially regulated, probably in response to long-term IOP exposure, in this animal model. The expression pattern of many genes is common to experimental models of elevated IOP and other retinal disorders such as diabetic retinopathy. However many genes are uniquely expressed in the retina of our model. This suggests that the mode of IOP elevation be it experimental or spontaneous could be relevant in determining which genes are regulated. Müller glia acquire a reactive phenotype as indicated by the upregulation of GFAP, c-myc, SA, and other Müller cell markers, emphasizing their relevance in pressure related- and other types of retinal injury. These data provide further evidence that IOP-mediated retinal injury is multifactorial and depends upon the interaction of different neuronal, glial, extracellular matrix, and vasogenic components.

Animals↗
PubMed2006

Cortisol promotes survival and regeneration of axotomised retinal ganglion cells and enhances effects of aurintricarboxylic acid.

BACKGROUND: Neuroprotection is essential for repair processes after a traumatic insult in the central nervous system. We have demonstrated previously significant neuroprotective properties of the anti-apoptotic drug aurintricarboxylic acid in the model of axotomised retinal ganglion cells. Glucocorticoids are widely used to treat injuries of the nervous system. Due to the anti-inflammatory and microglia-inhibiting properties of glucocorticoids, we studied the neuroprotective effects of intravitreally administered cortisol after an optic nerve cut. METHODS: Ninety-eight adult Sprague-Dawley rats were used in this study. The optic nerve was cut intra-orbitally. Either vehicle or compound solution was injected intravitreally. Fluorescent dye was put onto the optic nerve stump to label retinal ganglion cells retrogradely. Retinal whole mounts were prepared 2 weeks after axotomy, and surviving retinal ganglion cells were counted. RESULTS: Two weeks after axotomy, up to 50+/-7% of all retinal ganglion cells survived if cortisol was injected into the eye compared with 17+/-5% survival if only vehicle solution was injected. The neuroprotective effects of aurintricarboxylic acid (43+/-5% survival) could be further enhanced if combined with cortisol (up to 61+/-5% survival). Regeneration of cut retinal ganglion cell axons into a peripheral nerve graft could also be enhanced by an intravitreal injection of cortisol (169+/-42 regenerating retinal ganglion cells per mm2 vs. 73+/-12 cells per mm2 after vehicle injection). The increase was not as high as with aurintricarboxylic acid (192+/-40 cells per mm2), although more retinal ganglion cells survived with cortisol. This indicates that neuronal survival alone is not sufficient for subsequent axonal regeneration. Nevertheless, regeneration could be markedly increased if aurintricarboxylic acid and cortisol were combined (308+/-72 cells per mm2). CONCLUSIONS: Whereas aurintricarboxylic acid seems to act directly on lesioned retinal ganglion cells, cortisol seems to act on the glial environment, as indicated by microglial cell morphology and enhanced glial fibrillary acidic protein expression. The results show that both neuroprotection and regeneration can be enhanced by the combination of two simple compounds acting on different sites.

Animals↗
PubMed2006

Morphometric examination of human and monkey retinal ganglion cells within the papillomacular area.

PURPOSE: To examine the morphology of the retinal ganglion cells (RGCs) in the lesser characterized area lying between the optic disk and the macula that consists of the central papillomacular area (PMA) and the arcuate papillomacular bundle (PMB). METHODS: Nineteen human and 10 monkey (Macaca fascicularis) retinas obtained after death were used in the study. Perikaryal, axonal, and dendritic silhouettes were examined by postvital application of the fluorescent dye DiI, which specifically labeled RGCs when placed onto the optic fiber layer. The retinas were freed from surrounding tissue, prepared as flat mounts on a nitrocellulose filter, and fixed overnight in 4% paraformaldehyde. DiI diffuses along the membranes of ganglion cell axons, thereby completely labeling them, their cell bodies, and dendrites, which enables the RGCs to be examined with fluorescence microscopy. RESULTS: In both species, midget cells represented most of the RGCs within the PMA (96.15%) and possessed small, umbrella-like dendrites oriented toward the deeper retinal layers. Parasol cells were less abundant in both species and had small, typical symmetric dendrites. Also along the PMB, midget cells represented most cells (91.52%), whereas only 8.47% could be categorized as parasol cells. In both species, parasol cells of the PMB extended dendrites, which were oriented perpendicular to the axons. CONCLUSIONS: The data show that the PMA and PMB mainly contain small midget cells of typical morphology and size but with atypically oriented dendrites, which are only characteristic for this retinal area.

Aged↗
PubMed2005

Intraocular pressure changes during high-altitude acclimatization.

BACKGROUND: To evaluate the relationship between hypobaric hypoxia acclimatization and intraocular pressure (IOP) during ascent, acclimatization, and descent between 2286 m and 5050 m. METHODS: The following acclimatization-indicative physiological parameters were compared daily with IOP changes in eight healthy climbers of the 2003 Greek Karakorum expedition in altitude stages between 500 m and 5050 m: hemoglobin oxygen saturation (PO2), resting heart rate, blood pressure, retinal findings, and the Lake Louise score for acclimatization grading. RESULTS: IOP decreased significantly in the ascent phase (0.58 mmHg/100 m) and recovered (0.71 mmHg/100 m) during acclimatization and descent. A direct proportional correlation between decreases in PO2 and IOP was evaluated. Arterial blood pulse and pressure increased during acclimatization, while IOP decreased. No retinal hemorrhages were observed in well-acclimatized and incompletely acclimatized climbers. CONCLUSIONS: Every new active exposure to hypobaric hypoxia in the ascent phase induced a decrease in the IOP parallel to the PO2 decrease and to the level of acclimatization. The results from our study suggest that IOP changes are related to hypoxia-induced respiratory alkalosis and acclimatization stage, which could be used as a simple mobile screening test for acclimatization level to reveal acute mountain sickness and its severe consequences.

Acclimatization↗
PubMed2005

Lens epithelium supports axonal regeneration of retinal ganglion cells in a coculture model in vitro.

The purpose of this study was to determine whether the lens epithelium influences the survival or axonal growth of regenerating retinal ganglion cells. The optic nerves of adult albino rats were injured in order to induce axonal regeneration, and axon growth was then studied in retinal explants in the presence of cocultivated lens capsules carrying living epithelial cells. In the first series of experiments, cocultivation of retinal explants with lens epithelium in immediate proximity resulted in penetration of fibers into the lens epithelium, indicating that it supported axonal growth. In the second series of experiments, co-explants were placed 0.5-1.0mm from each other. The numbers of outgrowing retinal axons were determined both with respect to the retinal eccentricity and the topological relationship with the lenticular co-explant. The Wilcoxon matched-pairs signed-rank test was used to determine if the numbers of axons differed significantly between four regions of the explants. Significantly more axons grew out from the retinal edge facing the lenticular explant than from its opposite side, indicating that the lens epithelium supports axon growth. The numbers of surviving retinal ganglion cells in culture were determined after retrograde prelabelling with a neuroanatomical tracer. The number of fluorescent ganglion cells within the retinal explants did not significantly differ between the groups (Mann-Whitney test). These findings indicate that the lens epithelium influences both the amount of axonal regeneration and the direction of growth without affecting the survival rate of retinal ganglion cells in vitro.

Animals↗
PubMed2005

Recovery of visual evoked potentials after regeneration of cut retinal ganglion cell axons within the ascending visual pathway in adult rats.

PURPOSE: Following optic nerve damage, retinal ganglion cells (RGCs) fail to regenerate their axons and soon undergo apoptosis. However, many RGCs survive axotomy and regenerate lengthy axons after a lens injury (LI). If the cut optic nerve is re-sutured, RGC axons grow into the distal part of the optic nerve and reach their natural targets within the thalamus and midbrain. In this study, we check time-dependence and extent of restoration of flash visual evoked potentials (FVEPs) to examine the functional relevance of the regenerated retinogeniculate pathway. METHODS: The optic nerve in adult rats was cut and re-sutured. The lens was injured transsclerally using a pointed glass capillary. FVEPs were measured starting at the time point of surgery, and then repeatedly up to an age of several months. RESULTS: Detectable FVEPs appeared approximately ten weeks after the surgery, and their amplitudes increased during the next months to reach eventually 15-40% of their values before surgery. CONCLUSIONS: Partial restoration of FVEPs indicates that some regenerating RGC axons have "bridged" the distance between the eye and the central targets forming a functional re-connection of the corresponding RGC with thalamic target neurones to elicit recordable activation of the visual cortex.

Animals↗
PubMed2004

Decorin deficiency leads to impaired angiogenesis in injured mouse cornea.

Small leucine-rich proteoglycans play important roles in the organization of the extracellular matrix as well as for the regulation of cell behavior; two biological processes that are essential for angiogenesis. We investigated consequences of the targeted ablation of decorin (DCN), biglycan (BGN) and fibromodulin (FMOD) genes on inflammation-induced angiogenesis in the cornea. In wild-type mice, DCN was localized exclusively to the corneal stroma, while FMOD and BGN were more prominently expressed in epithelial cells. Endothelial cells from limbus blood vessels expressed BGN and FMOD, but no DCN. However, after induction of angiogenesis by chemical cauterization, DCN was expressed in the newly formed capillaries, together with BGN and FMOD. Notably, in DCN-deficient mice, the growth of vessels was significantly diminished, whereas it did not significantly change in FMOD- or BGN-deficient animals. Moreover, blood vessels of DCN-deficient mice exhibited a similar expression level of BGN as control mice, while FMOD was increased on day 3 after injury. These results indicate that DCN, in addition to its effects on fibrillogenesis, plays a regulatory role in angiogenesis and that FMOD in endothelial cells may be able to partially substitute for DCN.

Animals↗
PubMed2004

Switching mature retinal ganglion cells to a robust growth state in vivo: gene expression and synergy with RhoA inactivation.

The inability of mature CNS neurons to regenerate injured axons has been attributed to a loss of inherent growth potential of cells and to inhibitory signals associated with myelin and the glial scar. The present study investigated two complementary issues: (1) whether mature CNS neurons can be stimulated to alter their gene expression profile and switch into a strong growth state; and (2) whether inactivating RhoA, a convergence point for multiple inhibitory signals, is sufficient to produce strong regeneration even without activating the growth state of neurons. In the mature rat, retinal ganglion cells (RGCs) normally fail to regenerate axons through the injured optic nerve but can be stimulated to do so by activating macrophages in the eye (e.g., by lens injury). To investigate underlying changes in gene expression, we retrogradely labeled RGCs with a fluorescent dye, performed optic nerve surgery with or without lens injury, and 4 d later, dissociated retinas, isolated RGCs by fluorescence-activated cell sorting, and examined their profiles of gene expression using microarrays. To investigate the effects of inactivating RhoA, we transfected RGCs with adeno-associated viruses carrying a gene for C3 ribosyltransferase. Our results show that, with appropriate stimulation, mature CNS neurons can undergo dramatic changes in gene expression comparable with those seen in regenerating neurons of the PNS, and that RhoA inactivation by itself results in moderate regeneration, and strongly potentiates axon regeneration through the mature optic nerve when the growth state of neurons is activated.

ADP Ribose Transferases↗
PubMed2004

Ultrasonic visualization of the effect of blinking on the lacrimal pump mechanism.

BACKGROUND: The role of the lacrimal sac (LS) and the medial canthal tendon in the lacrimal pump mechanism is controversial. This study used ultrasonic visualization to analyze this phenomenon. METHODS: Movements of the LS and the medial canthal tendon during blinking were visualized with sonography. In addition, the maximal profile area of the LS was measured before and after blinking using 15-MHz sonography in 14 individuals with a normal lacrimal drainage system and in six patients with lacrimal duct obstruction. RESULTS: The upper part of the LS could be located as an echolucent structure between the lacrimal bone and the medial canthal tendon. The medial canthal tendon appeared to compress the LS during lid closure and release the LS during lid opening. The measured profile area of the visible normal LS at the compression time decreased by 50%. The dilated LS of patients with obstruction could also be compressed by the orbital muscle on blinking, but the maximum area decrease was only 15.5%. CONCLUSION: The findings imply that the lacrimal part of the orbicularis muscle contracts during blinking, with the medial canthal tendon compressing the LS in a cranial direction. Completion of lid closure then compresses both canaliculi and LS, forcing the intrasacral fluid through the drainage system. The expansion of the LS during the opening phase of the blink causes suction, and after opening of the punctal areas the canaliculi and LS vacuum breaks to reload with tear fluid. These findings demonstrate the importance of the orbicularis muscle and the medial canthal tendon for the lacrimal pump mechanism during blinking.

Adult↗
PubMed2004

Neuronal MAP2 mRNA: species-dependent differential dendritic targeting competence.

Providing the basis for local protein synthesis in dendritic microdomains, RNA transport in dendrites is thought to be underlying long-term neuronal plasticity. Dendritic RNA targeting mechanisms can therefore be expected to confer selective advantages in the evolution of complex neural systems. The question thus arises as to when and how dendritically targeted transcripts first acquired their targeting competence. To address this question, the dendritic targeting competence of MAP2 transcripts was examined in chicken, mouse and rat. In one approach, we established the somato-dendritic distribution of MAP2 transcripts in vivo. We found that in contrast to rodent MAP2 mRNAs, which are highly enriched in dendritic regions of the retina, chicken MAP2 transcripts are virtually absent from such areas and are rather confined to neuronal somata. In an independent line of investigation, we determined that a dendritic targeting element (DTE) corresponding to the mammalian MAP2 DTE is not contained in the 3' untranslated region (UTR) of avian MAP2 mRNA. The combined results indicate that in contrast to mammalian MAP2 transcripts, avian MAP2 mRNA is lacking dendritic targeting competence. The data thus suggest that the acquisition of such competence has likely been a relatively recent event in evolution.

Animals↗
PubMed2004

Distribution of scotoma pattern related to chiasmal lesions with special reference to anterior junction syndrome.

PURPOSE: To evaluate pathogenetic mechanisms and frequency distribution of visual field defects (VFDs) in patients with chiasmal lesions. Secondly, to reconsider the existence of "Wilbrand's knee" as far as referable to the anterior junction syndrome. METHODS: Consecutive visual field records related to chiasmal lesions were retrieved from the Tuebingen Perimetric Database. In all cases, at least one eye was examined with the Tuebingen Automated Perimeter using a standardized grid of 191 static targets within the central 30 degrees visual field, and a threshold-related, slightly supraliminal strategy. VFDs were classified according to standard neuro-ophthalmological categories. RESULTS: Results from 153 consecutive patients (65 male, 88 female) were evaluable. The majority (65%) of chiasmal lesions was due to pituitary adenoma, followed by craniopharyngioma (12%), astrocytoma (9%), and meningioma (8%). Vascular lesions in this region occurred rarely (2%). Three per cent of all patients had no final diagnosis. The majority (22%) of scotomas was attributable to involvement of the temporal hemifield in both eyes, with true bitemporal hemianopia being a very rare event (1%). Anterior junction syndrome, characterized by advanced visual field loss affecting the visual field centre in one eye and (possibly subtle) defects respecting the vertical midline in the fellow eye, was the second most frequent classifiable VFD (13%). Homonymous hemianopic VFDs occurred in 11% of all cases. Nine per cent of all patients exhibited monocular VFDs which did not respect the vertical midline, whereas in 3% of the subjects the monocular VFDs did not cross the vertical meridian. Binasal defects and posterior junction syndrome also occurred seldom (< 1%). Nineteen per cent of all visual field records of patients with chiasmal lesions had results, which could not be classified unequivocally, and an identical portion was rated normal. CONCLUSION: In patients with chiasmal lesions, incomplete involvement of the temporal hemifields in both eyes was the most frequent event (22%), followed by anterior junction syndrome (13%). The latter entity at least clinically indicates the proximity of the pre-chiasmal ipsilateral optic nerve and decussating fibres emanating from the inferior nasal hemiretina of the fellow eye. However, this cannot provide conclusive evidence for the existence of anterior Wilbrand's knee.

Adolescent↗
PubMed2004

Neuro-glial interactions in the adult rat retina after reaxotomy of ganglion cells: examination of neuron survival and phagocytic microglia using fluorescent tracers.

Retinal ganglion cells (RGCs) regenerating through peripheral nerve grafts show enhanced survival after further axonal injury for at least 4 weeks [Restor. Neurol. Neurosci. 21 (2003) 11]. Here, we examined the survival of the neurons and their microglial phagocytosis in dependence of the site of reaxotomy. Therefore, the optic nerve in adult rats was transected at different distances from the eye cup and replaced with an autologous piece of sciatic nerve. After 14 days of axonal growth, the regenerated neurites were reaxotomized either within the remaining optic stump or within the graft and their cell bodies were retrogradely labeled. Reaxotomy of regenerated ganglion cells within the remaining optic nerve resulted in reduced (but not significant) ganglion cell survival and significant microglial phagocytosis in contrast to reaxotomy within the peripheral nerve graft. Furthermore, phagocytosis-dependent labeling using two different fluorescent tracers revealed that the same microglial cell can phagocytose further dying ganglion cells within 14 days after the first activation. The results suggest that the intrasciatic segments of axons receive some trophic support that is retrogradely transported and required to limit the microglial activation. The microglial capability to phagocytose dying neurons several fold emphasizes their function in permanent scavenging within the retina.

Animals↗
PubMed2004

Potential role of Pax-2 in retinal axon navigation through the chick optic nerve stalk and optic chiasm.

The degree of fiber decussation at the optic chiasm differs between species, ranging from complete crossing in lower vertebrates to highly complex patterns of intermingling of the fibers from the two eyes seen in mammals and birds. Understanding the genetic control of fiber guidance through the chiasm is therefore important to unravel the developmental mechanisms within the visual system. Here we first report on early stages of chiasm formation, with pioneering axons from the left eye consistently arriving earlier than their counterparts from the right eye. This initial left-right asymmetry is transient and no functional significance is assigned to it yet. Secondly, we examined formation of the chiasm in relation with the expression of the transcription factor Pax-2 along the ventral eye cup and optic nerve stalk. Finally, in order to examine causal involvement of Pax-2 in chiasm formation, the gene was overexpressed along the neuraxis and in the eye cup at embryonic stages preceding the exit of axons from the eye, and hence arrival of axons at the chiasm. When studied with neuroanatomical tracing, Pax-2 overexpression resulted in visibly anomalous decussation of axons at the chiasm. A likely consequence of this perturbation was erroneous arrival of axons at the tectum, as observed by anterograde staining from the retina. These data suggest that balanced expression of Pax-2 results in the correct formation of the chick chiasm at early stages by imposing accurate pathfinding within the optic stalk and the midline.

Animals↗
PubMed2004

Presurgical and postsurgical ultrasound assessment of lacrimal drainage dysfunction.

PURPOSE: In addition to the obligatory clinical tests, imaging of the lacrimal drainage system (LDS) is useful in its clinical evaluation. The purpose of this study was to examine the usability and reliability of ultrasonography in the evaluation of the lacrimal drainage system. DESIGN: Observational cohort study. METHODS: A prospective study was conducted at a single institution. We performed ultrasound examinations on 17 patients with epiphora before and after surgery, and on 17 asymptomatic volunteers, to visualize and evaluate the anatomic and functional condition or pathologic abnormalities of the LDS. RESULTS: Echographic evaluation of the LDS was possible in all individuals. Pathologic abnormalities (canaliculitis, diverticulitis, concretion, or dilation of the lacrimal sac, and reduced functionality of the orbicular muscle and/or lacrimal sac pump) could be well demonstrated. In the postsurgical course, functional patency of the dacryocystorhinostomy opening could be verified in all cases. CONCLUSIONS: Sonography of the LDS appears to represent a reliable diagnostic technique supplementary to clinical tests in the presurgical and postsurgical examination of patients with epiphora. Pathologic abnormalities that may not be apparent in routine x-ray dacryocystography can be demonstrated with ultrasound techniques. Patients also benefit from the avoidance of exposure to ionizing radiation. However, ultrasound is not suitable for imaging the lower part of the lacrimal sac and the lacrimal duct because of the presence of overlying bony structures.

Adult↗
PubMed2004

Correlation between retinal ganglion cell death and chronically developing inherited glaucoma in a new rat mutant.

Glaucoma is a progressive optic neuropathy with characteristic optic disc changes, retinal ganglion cell loss and progressive visual field defects. Elevated intraocular pressure is considered to be a major risk factor in glaucomatous neuropathy. This study aimed to characterize and document a new chronic glaucoma model in the rat with respect to the effect of elevated intraocular pressure on overall retinal dysfunction and retinal ganglion cell loss, and to elucidate the possible mechanisms underlying this cell loss. Intraocular pressure (IOP) was measured in rats using a Tonopen. RGCs were retrogradely labeled with the fluorescent dye, 4-[didecylaminostyryl]-N-methyl-pyridinium-iodide (4-Di-10 ASP) and quantified on retinal flat mounts using fluorescence microscopy. The optic nerve head was examined fundoscopically. Changes in the histological appearance of the whole eyes was studied in paraffin sections, and immunohistochemistry was carried out on cryostat sections. The levels of mRNA for several genes were compared between control and glaucomatous retinae using semi-quantitative RT-PCR. Mutant animals are affected with either a unilateral or bilateral enlargement of the globes having an IOP that ranged from 25 to 45 mmHg, as compared to control values of 12-16 mmHg. The IOP of glaucomatous eyes increased significantly with age to attain a value of 35+/-7.3 at 1.5 years. Concomitant with the rise in IOP, the number of labeled RGCs continued to decrease in number with age. A total of 1887+/-117RGC mm(-2) could be labeled in wild-type control and juvenile mutant pre-glaucomatous retinas, whereas this number dropped to 92+/-26RGC mm(-2) at 1.5 years. Ophthalmoscopy revealed atrophied optic nerve heads in the affected eyes. The pars plicata and the pars plana of the ciliary body of glaucomatous eyes were hypertrophied and elongated, respectively. The anterior chamber was narrow and the irido-corneal angle open in glaucoma eyes. The mRNA of glial-fibrillary-acidic protein, endothelin-1, STAT-3 and STAT-6 increased in the retinas correlating with the severity and duration of the disease. Changes in the expression of GFAP and endothelin-1 could be confirmed using immunohistochemistry. This model may help to address several fundamental issues in the pathogenesis of glaucoma and aid in the development of neuroprotective strategies.

Animals↗